Figure 3
Effect of FasL and FADD expression in human glioma xenograft in vivo. Kaplan-Meier survival analysis of mice receiving (A) ΔGli36 cells preinfected with pG8-FasL and pG8-FADD, (B) intracranial injections of pG8-FasL and pG8-FADD into pre-established ΔGli36 tumor in comparison to controls (n = 5). PCR analysis performed on Hirt's DNA isolated from the mouse brain using primers against the exogenous eGFP gene and normalized using 18S. (C) Flow cytometry analysis of the percentage of eGFP+ cells in mice injected with 1 × 106 TU of (ii) pG8-18 amplicon vectors in comparison to (i) control. (iii), Photomicrograph of brain section showed the spreading of the pG8-18 viral vector. Image was captured using a 20×/N.A 0.50 Plan Fluor lens mounted on an Axioimager inverted microscope. r1 and r2 are the distance from the center of the eGFP+ region. The area of dispersion was calculated using the LSM Image browser (Zeiss) based on the formula for ellipse area = π*r1*r2. (D) PCR analysis performed on Hirt's DNA isolated from the non-tumor-bearing hemisphere (N) and the tumor-bearing hemisphere (T) of a mouse brain injected with pG8-18 amplicon viral vectors. PCR was performed using primers specific for luciferase and 18S. (E) Immunohistochemistry images showing the immunogenicity of pG8-18 amplicon viral vectors (1×106 TU) inoculated into immunocompetent Balb/c mice. Sections were stained for CD4, CD8 and CD11b on days 1 and 4 post-injection and counterstained with methyl green. Images shown are original magnification ×200.